• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of embojLink to Publisher's site
EMBO J. May 1993; 12(5): 2159–2168.
PMCID: PMC413437

Reduction of the disulfide bond of chromogranin B (secretogranin I) in the trans-Golgi network causes its missorting to the constitutive secretory pathways.

Abstract

The role of the single, highly conserved disulfide bond in chromogranin B (secretogranin I) on the sorting of this regulated secretory protein to secretory granules was investigated in the neuroendocrine cell line PC12. Treatment of PC12 cells with dithiothreitol (DTT), a membrane permeable thiol reducing agent known to prevent disulfide bond formation in intact cells, resulted in the secretion of newly synthesized chromogranin B, but only slightly decreased the intracellular storage of newly synthesized secretogranin II, a regulated secretory protein devoid of cysteines. The secretion of newly synthesized chromogranin B in the presence of DTT occurred with similar kinetics to those of a heparan sulfate proteoglycan, a known marker of the constitutive secretory pathway in PC12 cells. Analysis of the various secretory vesicles derived from the trans-Golgi network (TGN) indicated that DTT treatment diverted newly synthesized chromogranin B to constitutive secretory vesicles, whereas the packaging of secretogranin II into immature secretory granules was unaffected by the reducing agent. The chromogranin B molecules diverted to constitutive secretory vesicles, in contrast to those stored in secretory granules, were found to contain free sulfhydryl residues. The effect of DTT on chromogranin B occurred in the TGN rather than in the endoplasmic reticulum. We conclude that the sorting of CgB in the TGN to secretory granules is dependent upon the integrity of its single disulfide bond.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.1M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Alberini CM, Bet P, Milstein C, Sitia R. Secretion of immunoglobulin M assembly intermediates in the presence of reducing agents. Nature. 1990 Oct 4;347(6292):485–487. [PubMed]
  • Baeuerle PA, Huttner WB. Tyrosine sulfation is a trans-Golgi-specific protein modification. J Cell Biol. 1987 Dec;105(6 Pt 1):2655–2664. [PMC free article] [PubMed]
  • Benedum UM, Baeuerle PA, Konecki DS, Frank R, Powell J, Mallet J, Huttner WB. The primary structure of bovine chromogranin A: a representative of a class of acidic secretory proteins common to a variety of peptidergic cells. EMBO J. 1986 Jul;5(7):1495–1502. [PMC free article] [PubMed]
  • Benedum UM, Lamouroux A, Konecki DS, Rosa P, Hille A, Baeuerle PA, Frank R, Lottspeich F, Mallet J, Huttner WB. The primary structure of human secretogranin I (chromogranin B): comparison with chromogranin A reveals homologous terminal domains and a large intervening variable region. EMBO J. 1987 May;6(5):1203–1211. [PMC free article] [PubMed]
  • Braakman I, Helenius J, Helenius A. Manipulating disulfide bond formation and protein folding in the endoplasmic reticulum. EMBO J. 1992 May;11(5):1717–1722. [PMC free article] [PubMed]
  • Braakman I, Helenius J, Helenius A. Role of ATP and disulphide bonds during protein folding in the endoplasmic reticulum. Nature. 1992 Mar 19;356(6366):260–262. [PubMed]
  • Burgess TL, Kelly RB. Constitutive and regulated secretion of proteins. Annu Rev Cell Biol. 1987;3:243–293. [PubMed]
  • Chanat E, Huttner WB. Milieu-induced, selective aggregation of regulated secretory proteins in the trans-Golgi network. J Cell Biol. 1991 Dec;115(6):1505–1519. [PMC free article] [PubMed]
  • Freedman RB. Protein disulfide isomerase: multiple roles in the modification of nascent secretory proteins. Cell. 1989 Jun 30;57(7):1069–1072. [PubMed]
  • Friederich E, Fritz HJ, Huttner WB. Inhibition of tyrosine sulfation in the trans-Golgi retards the transport of a constitutively secreted protein to the cell surface. J Cell Biol. 1988 Nov;107(5):1655–1667. [PMC free article] [PubMed]
  • Gerdes HH, Rosa P, Phillips E, Baeuerle PA, Frank R, Argos P, Huttner WB. The primary structure of human secretogranin II, a widespread tyrosine-sulfated secretory granule protein that exhibits low pH- and calcium-induced aggregation. J Biol Chem. 1989 Jul 15;264(20):12009–12015. [PubMed]
  • Huttner WB, Gerdes HH, Rosa P. The granin (chromogranin/secretogranin) family. Trends Biochem Sci. 1991 Jan;16(1):27–30. [PubMed]
  • Iacangelo A, Affolter HU, Eiden LE, Herbert E, Grimes M. Bovine chromogranin A sequence and distribution of its messenger RNA in endocrine tissues. Nature. 1986 Sep 4;323(6083):82–86. [PubMed]
  • Iacangelo AL, Grimes M, Eiden LE. The bovine chromogranin A gene: structural basis for hormone regulation and generation of biologically active peptides. Mol Endocrinol. 1991 Nov;5(11):1651–1660. [PubMed]
  • Kimura JH, Lohmander LS, Hascall VC. Studies on the biosynthesis of cartilage proteoglycan in a model system of cultured chondrocytes from the Swarm rat chondrosarcoma. J Cell Biochem. 1984;26(4):261–278. [PubMed]
  • Lee RW, Huttner WB. Tyrosine-O-sulfated proteins of PC12 pheochromocytoma cells and their sulfation by a tyrosylprotein sulfotransferase. J Biol Chem. 1983 Sep 25;258(18):11326–11334. [PubMed]
  • Mayadas TN, Wagner DD. Vicinal cysteines in the prosequence play a role in von Willebrand factor multimer assembly. Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3531–3535. [PMC free article] [PubMed]
  • Peter F, Nguyen Van P, Söling HD. Different sorting of Lys-Asp-Glu-Leu proteins in rat liver. J Biol Chem. 1992 May 25;267(15):10631–10637. [PubMed]
  • Pimplikar SW, Huttner WB. Chromogranin B (secretogranin I), a secretory protein of the regulated pathway, is also present in a tightly membrane-associated form in PC12 cells. J Biol Chem. 1992 Feb 25;267(6):4110–4118. [PubMed]
  • Pohl TM, Phillips E, Song KY, Gerdes HH, Huttner WB, Rüther U. The organisation of the mouse chromogranin B (secretogranin I) gene. FEBS Lett. 1990 Mar 26;262(2):219–224. [PubMed]
  • Régnier-Vigouroux A, Tooze SA, Huttner WB. Newly synthesized synaptophysin is transported to synaptic-like microvesicles via constitutive secretory vesicles and the plasma membrane. EMBO J. 1991 Dec;10(12):3589–3601. [PMC free article] [PubMed]
  • Rosa P, Fumagalli G, Zanini A, Huttner WB. The major tyrosine-sulfated protein of the bovine anterior pituitary is a secretory protein present in gonadotrophs, thyrotrophs, mammotrophs, and corticotrophs. J Cell Biol. 1985 Mar;100(3):928–937. [PMC free article] [PubMed]
  • Rosa P, Hille A, Lee RW, Zanini A, De Camilli P, Huttner WB. Secretogranins I and II: two tyrosine-sulfated secretory proteins common to a variety of cells secreting peptides by the regulated pathway. J Cell Biol. 1985 Nov;101(5 Pt 1):1999–2011. [PMC free article] [PubMed]
  • Rosa P, Mantovani S, Rosboch R, Huttner WB. Monensin and brefeldin A differentially affect the phosphorylation and sulfation of secretory proteins. J Biol Chem. 1992 Jun 15;267(17):12227–12232. [PubMed]
  • Roy P, Chevrier D, Fournier H, Racine C, Zollinger M, Crine P, Boileau G. Investigation of a possible role of the amino-terminal pro-region of proopiomelanocortin in its processing and targeting to secretory granules. Mol Cell Endocrinol. 1991 Dec;82(2-3):237–250. [PubMed]
  • Schimmel A, Bräunling O, Rüther U, Huttner WB, Gerdes HH. The organisation of the mouse secretogranin II gene. FEBS Lett. 1992 Dec 21;314(3):375–380. [PubMed]
  • Simon JP, Aunis D. Biochemistry of the chromogranin A protein family. Biochem J. 1989 Aug 15;262(1):1–13. [PMC free article] [PubMed]
  • Tatu U, Braakman I, Helenius A. Membrane glycoprotein folding, oligomerization and intracellular transport: effects of dithiothreitol in living cells. EMBO J. 1993 May;12(5):2151–2157. [PMC free article] [PubMed]
  • Tooze SA, Huttner WB. Cell-free protein sorting to the regulated and constitutive secretory pathways. Cell. 1990 Mar 9;60(5):837–847. [PubMed]
  • Tooze SA, Huttner WB. Cell-free formation of immature secretory granules and constitutive secretory vesicles from trans-Golgi network. Methods Enzymol. 1992;219:81–93. [PubMed]
  • Tooze SA, Flatmark T, Tooze J, Huttner WB. Characterization of the immature secretory granule, an intermediate in granule biogenesis. J Cell Biol. 1991 Dec;115(6):1491–1503. [PMC free article] [PubMed]
  • Wagner DD. Cell biology of von Willebrand factor. Annu Rev Cell Biol. 1990;6:217–246. [PubMed]
  • Wu HJ, Rozansky DJ, Parmer RJ, Gill BM, O'Connor DT. Structure and function of the chromogranin A gene. Clues to evolution and tissue-specific expression. J Biol Chem. 1991 Jul 15;266(20):13130–13134. [PubMed]

Articles from The EMBO Journal are provided here courtesy of The European Molecular Biology Organization

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • Compound
    Compound
    PubChem Compound links
  • MedGen
    MedGen
    Related information in MedGen
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...